Institute of Transfusion Medicine and Immunohematology, German Red Cross, Johann Wolfgang Goethe University, Frankfurt, Germany.
Transfusion. 2009 Dec;49(12):2604-11. doi: 10.1111/j.1537-2995.2009.02326.x. Epub 2009 Jul 22.
According to German regulations and guidelines, residual red blood cells (rRBCs) and residual white blood cells (rWBCs) must number fewer than 3 x 10(9) cells/unit and 1 x 10(6) cells/unit in platelet concentrates (PCs), respectively. Due to low levels of residual cells in final products, there is still a need for fast, reliable, and sensitive methods of automated detection of these cell types.
In Part A, 21 PCs were spiked with predetermined numbers of red blood cells (RBCs) and white blood cells (WBCs). The linearity, precision, and accuracy of the BD Thrombo Count assay (BD Biosciences Europe) were tested and validated according to international guidelines. Finally in Part B, 100 PCs prepared from pooled buffy coats were tested by the BD Thrombo Count assay and compared with other methods, including Nageotte (rWBCs) and Neubauer (rRBCs) counting chambers and the flow cytometric BD LeucoCOUNT (Becton Dickinson) assay (rWBCs).
The unspecific background of blank PC samples was fewer than 0.02 cells/microL for WBCs and fewer than 34 cells/microL for RBCs (mean, 21). Linear regression and precision analyses of spiked PC samples were determined for both WBCs (r(2) = 0.992; range, 0.6-6.0 WBCs/microL) and RBCs (r(2) = 0.999; 800-8000 RBCs/microL). No carryover of cells or drift in results was detected in the automated sample acquisition mode. Analysis according to statistical methods of Bland and Altman demonstrated a high correlation between BD Thrombo Count and the Neubauer manual counting chamber.
This novel flow cytometric test is a quick and reliable single-tube assay that has been demonstrated as a potential alternative for the existing manual microscopic counting procedures that are both time-consuming and laborious.
根据德国法规和指南，血小板浓缩物（PCs）中残留的红细胞（rRBC）和白细胞（rWBC）数量应分别少于 3 x 10(9) 个/单位和 1 x 10(6) 个/单位。由于最终产品中残留细胞数量低，仍然需要快速、可靠和敏感的自动检测这些细胞类型的方法。
在 A 部分中，将 21 个 PCs 与预定数量的红细胞（RBC）和白细胞（WBC）混合。根据国际指南，测试和验证了 BD Thrombo Count 测定法（BD Biosciences Europe）的线性、精密度和准确性。最后，在 B 部分，使用 BD Thrombo Count 测定法测试了从混合 buffy coat 制备的 100 个 PCs，并与其他方法进行了比较，包括 Nageotte（rWBC）和 Neubauer（rRBC）计数室以及流式细胞术 BD LeucoCOUNT（Becton Dickinson）测定法（rWBC）。
空白 PC 样本的非特异性背景中，WBC 的数量少于 0.02 个/微升，RBC 的数量少于 34 个/微升（平均值，21）。对两种 WBC（r(2) = 0.992；范围，0.6-6.0 WBCs/microL）和 RBC（r(2) = 0.999；800-8000 RBCs/microL）的 PC 样本进行了线性回归和精密度分析。在自动样品采集模式下，未检测到细胞的携带或结果的漂移。根据 Bland 和 Altman 的统计方法分析表明，BD Thrombo Count 与 Neubauer 手动计数室高度相关。
这种新型流式细胞术检测是一种快速可靠的单管检测方法，已被证明是现有手动显微镜计数程序的潜在替代方法，这些程序既耗时又费力。
